Effect of rhVEGF gene transfection on survival of grafts after autologous free granular fat transplantation in rats

Objective： To investigate the effect of recombinant human vascular endothelial growth factor （rhVEGF） on autologous free granular fat grafts in rats.
Methods： Forty-eight Sprague Dawley （ SD ） rats, weighing 190-280 g and regardless sex, were randomly divided into three groups, sixteen in each. After fat transplantation, the rats were treated with plasmid DNA encoding rhVEGF protein （the experimental group ）, plasmid DNA （ the negative group） and normal saline （ the blank control group ）, respectively. At 3, 7, 15 and 30 days after transplantation, the rats were killed and the grafts were weighed, respectively. Histopathological changes were evaluated. Microvessel density and the expression of VEGF were examined by immunohistochemical staining and Western blotting.
Results： The weights of the negative and blank control groups were significantly reduced on the 7th, 15th and 30th days compared with those of the experimental group. The expression of VEGF and the microvessel density in the experimental group were significantly higher than the other two groups during the latter periods.
Conclusion： The plasmid encoding VEGF can induce expression of VEGF and angiogenesis in fat grafts and reduce the absorption of free fat grafts.     

大鼠部分肝缺血再灌注损伤后切除对肝再生的影响

目的 观察大鼠部分肝缺血再灌注损伤后切除对残肝再生的影响.方法 将75只健康雄性SD大鼠随机分为5组:肝脏左叶和中叶(约占全肝70%)切除组(Control组)、肝脏左叶和中叶缺血10min再灌注30min后切除组(I10R30组)、类推得到I60R30组、I90R30组、I90R60组.术后6、12、24h等时间点,测定再生肝重量(RLW);自动生化分析仪检测血清丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST)含量;酶联免疫吸附试验(ELISA)检测血清肿瘤坏死因子(TNF)-α含量;通过免疫组织化学法检测残肝增殖细胞核抗原(Ki-67)表达.结果 术后12h,I60R30、I90R30和I90R60组RLW值分别为(1.80±0.03)%、(1.82±0.10)%、(1.87±0.05)%;Ki-67值分别为(58.35±2.18)%、(59.73±3.06)%、(62.65±2.24)%,均明显高于对照组(P<0.05).缺血再灌注干预各组ALT和AST明显高于对照组(P<0.05).术后6h和12h,I60R30、I90R30和I90R60组TNF-α明显高于对照组(P<0.05).结论 大鼠即将被切除的肝脏先缺血再灌注后切除,对残肝再生具有促进作用;诱导产生的TNF-α表达量增多是促进肝再生的原因之一.

Abstract：

Objective To investigate the effects of ischemia reperfusion injury before partial hepatectomy on liver regeneration in rats. Methods Seventy-five male healthy SD rats were randomly classified into 5 groups: group control, in which rats were only subjected to 70% hepatectomy; group I10R30, 70% liver hepatectomy after 10 min of ischemia and 30 min of reperfusion in the resected liver; By analogy, group I60R30, group I90R30 and group I90R60 were constructed. At 6th, 12th and 24th h after operation, RLW was determined; serum alanine aminotransferase (ALT) and aspartate transaminase (AST) activities were measured by using autoanalyzer; the levels of serum tumor necrosis factor (TNF)-α were determined by ELISA and the expression level of Ki-67 was detected by using immunohistochemical methods in the residual liver tissues. Results At 12th h after partial hepatectomy, the rate of RLW in group I60R30, group I90R30 and group I90R60 was (1.80±0.03)%, (1.82±0.10)%, (1.87±0.05)% respectively; the rate of Ki-67 was (58.35±2.18)%, (59.73±3.06)%, (62.65±2.24)% respectively, which was significantly higher than that in the group control (P<0.05). The levels of ALT and AST in rats with ischemia reperfusion injury were higher than in the group control (P<0.05). At 6th h and 12th h after operation, the expression levels of TNF-α in groups I60R30, I90R30 and I90R60 were significantly higher than those in the group control (P<0.05). Conclusion Ischemia reperfusion injury in the resected liver before partial hepatectomy could improve liver regeneration of the remnant liver in rats. The high expression of induced TNF-α may be one of the reasons.     

不同剂量瑞芬太尼对大鼠肾脏缺血再灌注损伤的影响

目的 探讨不同剂量瑞芬太尼对大鼠肾脏缺血再灌注损伤的影响.方法 健康成年雄性SD大鼠60只,体重220～250 g,采用随机数字表法,将大鼠随机分为5组(n=12):假手术组(S组)、模型组(M组)、低、中和高剂量瑞芬太尼组(RL组、RM组和RH组).除S组外均采用夹闭双侧肾动脉45 min后恢复再灌注法建立肾脏缺血再灌注模型.RL组、RM组和RH组于缺血前15min分别经尾静脉输注瑞芬太尼0.2、0.6、1.0μg·kg-1·min-1至再灌注30 min;S组和M组给予等容量生理盐水替代.于再灌注30 min及24 h时经股静脉采集血样1 ml,测定血清BUN及Cr浓度;于再灌注24h时取肾组织,测定MDA含量及SOD、Ca2+-ATP酶活性,光、电镜下观察肾组织病理学结果.结果 与S组比较,其余4组血清BUN和Cr浓度、肾组织MDA含量升高,SOD和Ca2+-ATP酶活性降低(P＜0.05或0.01),肾组织有不同程度的病理学损伤.与M组比较,RL组、RM组和RH组血清BUN和Cr浓度、肾组织MDA含量降低,SOD和Ca2+-ATP酶活性升高(P＜0.05或0.01),肾组织病理学损伤减轻o RL组、RM组和RH组随瑞芬太尼剂量增加,血清BUN和Cr浓度、肾组织MDA含量逐渐降低,SOD和Ca2+-ATP酶活性逐渐升高(P＜0.05或0.01),肾组织病理学损伤逐渐减轻.结论 瑞芬太尼可减轻大鼠肾脏缺血再灌注损伤,且与剂量有关,其机制与抑制脂质过氧化反应、提高Ca2+-ATP酶活性有关.

Abstract：

Objective To investigate the effects of different doses of remifentanil on the renal ischemiareperfusion (I/R) injury in rats. Methods Sixty male SD rats weighing 220-250 g were randomly divided into 5 groups ( n = 12 each): sham operation group (group S), model group (group M), low, median and high doses of remifentanil groups (RL, RM and RH groups). The rats were anesthetized with intraperitoneal 5% chloral hydrate 6 ml/kg. Renal ischemia was induced by clamping the bilateral renal arteries for 45 min using an atraumatwere infused via the caudal vein 15 min before ischemia respectively and the infusion was stopped at 30 min of reperfusion, while S and M groups received equal volume of normal saline instead. Blood samples were collected from the femoral vein at 30 min and 24 h of reperfusion for measurement of serum creatinine (Cr) and blood urea nitrogen (BUN) concentrations. The rats were sacrificed at 24 h of reperfusion and the renal tissues were removed for determination of MDA content, SOD and Ca2+ -ATPase activities. Pathological changes in renal tissues were observed with light and electron microscopes. Results Compared with group S, the concentrations of serum Cr and BUN and content of MDA were significantly increased, while activities of SOD and Ca2+ -ATPase were significantly decreased in the other 4 groups ( P ＜ 0.05 or 0.01). Compared with group M, the concentrations of serum Cr and BUN and content of MDA were significantly decreased, activities of SOD and Ca2+ -ATPase were significantly increased (P ＜0.05 or 0.01) and the pathological changes were reduced in RH, RM and RL groups. The plasma BUN and Cr concentrations and MDA content were decreased gradually and SOD and Ca2+ -ATPase activities were increased gradually with the increase in the doses of remifentanil in RL, RM and RH groups ( P ＜ 0.05 or 0.01 ).Remifentanil infusion significantly attenuated the pathologic changes in a dose-dependent manner. Conclusion Remifentanil can reduce the renal I/R injury in a dose-dependent manner by inhibiting lipid peroxidation and increasing Ca2+ -ATPase activity.     

吸入一氧化碳对大鼠脑死亡致肺损伤的影响

Objective To investigate the effects of carbon monoxide (CO) inhalation on lung injury induced by brain death (BD) in rats. Methods Adult male Wistar rats weighing 250-300 g were used in this study. The animals were anesthetized with intraperitoneal pentobarbital sodium 60 mg/kg, tracheostomized and mechanically ventilated (VT 10 ml/kg, RR 50 bpm, PEEP 2 cm H2O). A balloon-tip catheter was placed in the cranium. Twenty-four rats in which Fogarty catheter was successfully placed in the cranium without complication were randomly divided into 3 groups ( n = 8 each) : group I sham operation (group S) ; group II BD and group Ⅲ BDCO. BD was induced by increase in intracranial pressure produced by inflating the balloon at the tip of the catheter. In group S the balloon of the catheter was not inflated. The animals inhaled 40% O2 for 150 min. In group BD, BD was induced and confirmed at 30 min after inflation of the balloon. Then 40% O2 was inhaled for 120 min. In group BDCO, 40% O2 and 0.025% CO were inhaled for 120 min after BD was confirmed at 30 min after balloon inflation. At the end of the experiment the animals were killed. Arterial blood samples were obtained for blood gas analysis before anesthesia (basline), immediately after confirmation of BD, and at 30, 60, 90 and 120 min of CO inhalation. Blood was collected for determination of plasma TNF-α, IL-6 and IL-10 concentrations at 120 min of CO inhalation. The lungs were obtained for determination of W/D lung weight ratio, and MPO activity in the lung tissue and microscopic examination. Lung injury scores were calculated. Results PaO2/FiO2 was stable during the 150 min in group S. Brain death significantly decreased PaO2/FiO2 at 30 min after balloon inflation. PaO2/FiO2 was gradually decreasing during the 120 min in group BD. CO inhalation prevented PaO2/FiO2 from decreasing further. W/D lung weight ratio and MPO activity were significantly higher in group BD than in group S and BDCO. The lung injury score (1 = normal, 4= severely injured) and plasma TNF-αα IL-6 and IL-10 concentrations were significantly higher in group BD than in group S. CO inhalation ameliorated the BD-induced lung injury and attenuated the increase in plasma TNF-a and IL-6 concentration. Plasma IL-10 concentration was significantly higher in group BDCO than in group BD. Conclusion CO inhalation can ameliorate acute lung injury induced by BD through decreasing the local and systemic inflammatory response.     

吸入一氧化碳对大鼠脑死亡致肺损伤的影响

Objective To investigate the effects of carbon monoxide (CO) inhalation on lung injury induced by brain death (BD) in rats. Methods Adult male Wistar rats weighing 250-300 g were used in this study. The animals were anesthetized with intraperitoneal pentobarbital sodium 60 mg/kg, tracheostomized and mechanically ventilated (VT 10 ml/kg, RR 50 bpm, PEEP 2 cm H2O). A balloon-tip catheter was placed in the cranium. Twenty-four rats in which Fogarty catheter was successfully placed in the cranium without complication were randomly divided into 3 groups ( n = 8 each) : group I sham operation (group S) ; group II BD and group Ⅲ BDCO. BD was induced by increase in intracranial pressure produced by inflating the balloon at the tip of the catheter. In group S the balloon of the catheter was not inflated. The animals inhaled 40% O2 for 150 min. In group BD, BD was induced and confirmed at 30 min after inflation of the balloon. Then 40% O2 was inhaled for 120 min. In group BDCO, 40% O2 and 0.025% CO were inhaled for 120 min after BD was confirmed at 30 min after balloon inflation. At the end of the experiment the animals were killed. Arterial blood samples were obtained for blood gas analysis before anesthesia (basline), immediately after confirmation of BD, and at 30, 60, 90 and 120 min of CO inhalation. Blood was collected for determination of plasma TNF-α, IL-6 and IL-10 concentrations at 120 min of CO inhalation. The lungs were obtained for determination of W/D lung weight ratio, and MPO activity in the lung tissue and microscopic examination. Lung injury scores were calculated. Results PaO2/FiO2 was stable during the 150 min in group S. Brain death significantly decreased PaO2/FiO2 at 30 min after balloon inflation. PaO2/FiO2 was gradually decreasing during the 120 min in group BD. CO inhalation prevented PaO2/FiO2 from decreasing further. W/D lung weight ratio and MPO activity were significantly higher in group BD than in group S and BDCO. The lung injury score (1 = normal, 4= severely injured) and plasma TNF-αα IL-6 and IL-10 concentrations were significantly higher in group BD than in group S. CO inhalation ameliorated the BD-induced lung injury and attenuated the increase in plasma TNF-a and IL-6 concentration. Plasma IL-10 concentration was significantly higher in group BDCO than in group BD. Conclusion CO inhalation can ameliorate acute lung injury induced by BD through decreasing the local and systemic inflammatory response.     

缺血预处理-后处理对大鼠肠缺血再灌注损伤的影响

Objective To evaluate the effects of ischemic preconditioning-postconditioning on the intestinal ischemia-reperfusion (IR) injury in rats. Methods Forty healthy male SD weighing 225-275 g were randomly assigned into 5 groups ( n = 8 each): group I sham operation (group S) ; group II intestinal IR (group IIR); group Ⅲ ischemic preconditioning (group Ipr); group IV ischemic postconditioning (group Ipo); group V Ipr+ Ipo. The rats were anesthetized with intraperitonel 20% urethane 5 ml/kg. Superior mesenteric artery (SMA) was occluded for 60 min followed by 60 min reperfusion. In group S, SMA was isolated but not occluded. In group Ipr, SMA was occluded for 10 min followed by 10 min reperfusion, and the rest procedures were performed using the method described in group IIR. In group Ipo, 60 min ischemia was followed by three 30 s episodes of ischemia at 30 s intervals for reperfusion. In group Ipr+ Ipo, Ipr was performed followed by Ipo and the procedures were performed using the methods described in group Ipr and Ipo. The animals were killed at 60 min of reperfusion. The intestinal tissues were immediately removed for determination of MDA content, SOD and MPO activities and the degree of damage to intestinal mucous membrane was scored according to Chiu score. Arterial blood samples were taken for determination of plasma concentrations of TNF-α and 1L-6. Results Compared with group S, Chiu score, MDA content, MPO activity, and plasma concentrations of TNF-α and IL-6 were significantly increased, whereas SOD activity decreased in the other 4 groups ( P < 0.05). Chiu score, MDA content, MPO activity, and plasma concentrations of TNF-α and IL-6 were significantly decreased, whereas SOD activity increased in group Ipr, Ipo and Ipr + Ipo as compared with group IIR ( P < 0.05). Chiu score and MDA content were significantly lower, whereas SOD activity higher in group Ipr + Ipo than in group Ipr and Ipo ( P < 0.05). No significant differences were detected in the indices between group Ipr and group Ipo ( P > 0.05). Conclusion Ischemic preconditioning-postconditioning can attenuate the intestinal IR injury in rats, and the efficacy is better than that of either Ipr or Ipo alone.     

缺血预处理-后处理对大鼠肠缺血再灌注损伤的影响

Objective To evaluate the effects of ischemic preconditioning-postconditioning on the intestinal ischemia-reperfusion (IR) injury in rats. Methods Forty healthy male SD weighing 225-275 g were randomly assigned into 5 groups ( n = 8 each): group I sham operation (group S) ; group II intestinal IR (group IIR); group Ⅲ ischemic preconditioning (group Ipr); group IV ischemic postconditioning (group Ipo); group V Ipr+ Ipo. The rats were anesthetized with intraperitonel 20% urethane 5 ml/kg. Superior mesenteric artery (SMA) was occluded for 60 min followed by 60 min reperfusion. In group S, SMA was isolated but not occluded. In group Ipr, SMA was occluded for 10 min followed by 10 min reperfusion, and the rest procedures were performed using the method described in group IIR. In group Ipo, 60 min ischemia was followed by three 30 s episodes of ischemia at 30 s intervals for reperfusion. In group Ipr+ Ipo, Ipr was performed followed by Ipo and the procedures were performed using the methods described in group Ipr and Ipo. The animals were killed at 60 min of reperfusion. The intestinal tissues were immediately removed for determination of MDA content, SOD and MPO activities and the degree of damage to intestinal mucous membrane was scored according to Chiu score. Arterial blood samples were taken for determination of plasma concentrations of TNF-α and 1L-6. Results Compared with group S, Chiu score, MDA content, MPO activity, and plasma concentrations of TNF-α and IL-6 were significantly increased, whereas SOD activity decreased in the other 4 groups ( P < 0.05). Chiu score, MDA content, MPO activity, and plasma concentrations of TNF-α and IL-6 were significantly decreased, whereas SOD activity increased in group Ipr, Ipo and Ipr + Ipo as compared with group IIR ( P < 0.05). Chiu score and MDA content were significantly lower, whereas SOD activity higher in group Ipr + Ipo than in group Ipr and Ipo ( P < 0.05). No significant differences were detected in the indices between group Ipr and group Ipo ( P > 0.05). Conclusion Ischemic preconditioning-postconditioning can attenuate the intestinal IR injury in rats, and the efficacy is better than that of either Ipr or Ipo alone.     

吸入一氧化碳对大鼠脑死亡致肺损伤的影响

Objective To investigate the effects of carbon monoxide (CO) inhalation on lung injury induced by brain death (BD) in rats. Methods Adult male Wistar rats weighing 250-300 g were used in this study. The animals were anesthetized with intraperitoneal pentobarbital sodium 60 mg/kg, tracheostomized and mechanically ventilated (VT 10 ml/kg, RR 50 bpm, PEEP 2 cm H2O). A balloon-tip catheter was placed in the cranium. Twenty-four rats in which Fogarty catheter was successfully placed in the cranium without complication were randomly divided into 3 groups ( n = 8 each) : group I sham operation (group S) ; group II BD and group Ⅲ BDCO. BD was induced by increase in intracranial pressure produced by inflating the balloon at the tip of the catheter. In group S the balloon of the catheter was not inflated. The animals inhaled 40% O2 for 150 min. In group BD, BD was induced and confirmed at 30 min after inflation of the balloon. Then 40% O2 was inhaled for 120 min. In group BDCO, 40% O2 and 0.025% CO were inhaled for 120 min after BD was confirmed at 30 min after balloon inflation. At the end of the experiment the animals were killed. Arterial blood samples were obtained for blood gas analysis before anesthesia (basline), immediately after confirmation of BD, and at 30, 60, 90 and 120 min of CO inhalation. Blood was collected for determination of plasma TNF-α, IL-6 and IL-10 concentrations at 120 min of CO inhalation. The lungs were obtained for determination of W/D lung weight ratio, and MPO activity in the lung tissue and microscopic examination. Lung injury scores were calculated. Results PaO2/FiO2 was stable during the 150 min in group S. Brain death significantly decreased PaO2/FiO2 at 30 min after balloon inflation. PaO2/FiO2 was gradually decreasing during the 120 min in group BD. CO inhalation prevented PaO2/FiO2 from decreasing further. W/D lung weight ratio and MPO activity were significantly higher in group BD than in group S and BDCO. The lung injury score (1 = normal, 4= severely injured) and plasma TNF-αα IL-6 and IL-10 concentrations were significantly higher in group BD than in group S. CO inhalation ameliorated the BD-induced lung injury and attenuated the increase in plasma TNF-a and IL-6 concentration. Plasma IL-10 concentration was significantly higher in group BDCO than in group BD. Conclusion CO inhalation can ameliorate acute lung injury induced by BD through decreasing the local and systemic inflammatory response.     

吸入一氧化碳对大鼠脑死亡致肺损伤的影响

Objective To investigate the effects of carbon monoxide (CO) inhalation on lung injury induced by brain death (BD) in rats. Methods Adult male Wistar rats weighing 250-300 g were used in this study. The animals were anesthetized with intraperitoneal pentobarbital sodium 60 mg/kg, tracheostomized and mechanically ventilated (VT 10 ml/kg, RR 50 bpm, PEEP 2 cm H2O). A balloon-tip catheter was placed in the cranium. Twenty-four rats in which Fogarty catheter was successfully placed in the cranium without complication were randomly divided into 3 groups ( n = 8 each) : group I sham operation (group S) ; group II BD and group Ⅲ BDCO. BD was induced by increase in intracranial pressure produced by inflating the balloon at the tip of the catheter. In group S the balloon of the catheter was not inflated. The animals inhaled 40% O2 for 150 min. In group BD, BD was induced and confirmed at 30 min after inflation of the balloon. Then 40% O2 was inhaled for 120 min. In group BDCO, 40% O2 and 0.025% CO were inhaled for 120 min after BD was confirmed at 30 min after balloon inflation. At the end of the experiment the animals were killed. Arterial blood samples were obtained for blood gas analysis before anesthesia (basline), immediately after confirmation of BD, and at 30, 60, 90 and 120 min of CO inhalation. Blood was collected for determination of plasma TNF-α, IL-6 and IL-10 concentrations at 120 min of CO inhalation. The lungs were obtained for determination of W/D lung weight ratio, and MPO activity in the lung tissue and microscopic examination. Lung injury scores were calculated. Results PaO2/FiO2 was stable during the 150 min in group S. Brain death significantly decreased PaO2/FiO2 at 30 min after balloon inflation. PaO2/FiO2 was gradually decreasing during the 120 min in group BD. CO inhalation prevented PaO2/FiO2 from decreasing further. W/D lung weight ratio and MPO activity were significantly higher in group BD than in group S and BDCO. The lung injury score (1 = normal, 4= severely injured) and plasma TNF-αα IL-6 and IL-10 concentrations were significantly higher in group BD than in group S. CO inhalation ameliorated the BD-induced lung injury and attenuated the increase in plasma TNF-a and IL-6 concentration. Plasma IL-10 concentration was significantly higher in group BDCO than in group BD. Conclusion CO inhalation can ameliorate acute lung injury induced by BD through decreasing the local and systemic inflammatory response.     

吸入一氧化碳对大鼠脑死亡致肺损伤的影响

Objective To investigate the effects of carbon monoxide (CO) inhalation on lung injury induced by brain death (BD) in rats. Methods Adult male Wistar rats weighing 250-300 g were used in this study. The animals were anesthetized with intraperitoneal pentobarbital sodium 60 mg/kg, tracheostomized and mechanically ventilated (VT 10 ml/kg, RR 50 bpm, PEEP 2 cm H2O). A balloon-tip catheter was placed in the cranium. Twenty-four rats in which Fogarty catheter was successfully placed in the cranium without complication were randomly divided into 3 groups ( n = 8 each) : group I sham operation (group S) ; group II BD and group Ⅲ BDCO. BD was induced by increase in intracranial pressure produced by inflating the balloon at the tip of the catheter. In group S the balloon of the catheter was not inflated. The animals inhaled 40% O2 for 150 min. In group BD, BD was induced and confirmed at 30 min after inflation of the balloon. Then 40% O2 was inhaled for 120 min. In group BDCO, 40% O2 and 0.025% CO were inhaled for 120 min after BD was confirmed at 30 min after balloon inflation. At the end of the experiment the animals were killed. Arterial blood samples were obtained for blood gas analysis before anesthesia (basline), immediately after confirmation of BD, and at 30, 60, 90 and 120 min of CO inhalation. Blood was collected for determination of plasma TNF-α, IL-6 and IL-10 concentrations at 120 min of CO inhalation. The lungs were obtained for determination of W/D lung weight ratio, and MPO activity in the lung tissue and microscopic examination. Lung injury scores were calculated. Results PaO2/FiO2 was stable during the 150 min in group S. Brain death significantly decreased PaO2/FiO2 at 30 min after balloon inflation. PaO2/FiO2 was gradually decreasing during the 120 min in group BD. CO inhalation prevented PaO2/FiO2 from decreasing further. W/D lung weight ratio and MPO activity were significantly higher in group BD than in group S and BDCO. The lung injury score (1 = normal, 4= severely injured) and plasma TNF-αα IL-6 and IL-10 concentrations were significantly higher in group BD than in group S. CO inhalation ameliorated the BD-induced lung injury and attenuated the increase in plasma TNF-a and IL-6 concentration. Plasma IL-10 concentration was significantly higher in group BDCO than in group BD. Conclusion CO inhalation can ameliorate acute lung injury induced by BD through decreasing the local and systemic inflammatory response.     

Acceptable warm ischemia time of tracheal grafts from non-heart-beating donors in rats

Objectives

Warm ischemia (WI)-induced airway complications are common in clinical lung transplantation. However, the acceptable WI time of tracheal grafts from non–heart-beating donors (NHBDs) is unknown. The purpose of this study was to determine the acceptable WI time by observing tracheal epithelial regeneration among NHBD.

Method

Forty-eight rats were randomly divided into four groups (each with 12 rats): WI-0 minutes (group A), WI-30 minutes (group B), WI-45 minutes (group C), and WI-60 minutes (group D). In each group, the tracheas from 6 rats were imbedded in the greater omentum of 6 other rats. Fourteen days later, the transplanted trachea was obtained from the recipient to evaluate epithelial thickness and regeneration. Six tracheas were obtained from living donors as a control group.

Results

There were no significant differences in tracheal transplantation time (mean, 17.66 ± 1.21 minutes). There were no significant differences in epithelial thickness and regeneration between the controls and groups A, B, and C (P < .05). Group D showed no normal epithelial structure of the trachea only with monolayer cells.

Conclusions

The time limits of tolerance to WI of tracheal grafts from NHBDs may be 45 minutes.     

大鼠小体积肝移植后肝细胞再生的研究

目的 探讨小体积肝移植术后肝再生的情况。方法 建立大鼠30％原位肝移植模型,实验分为肝切除（PH）组、全肝移植（0LT）组和30％小体积肝移植（30％POLT）组,观察1w生存率,并于术后1、2、3、7d检测肝细胞增殖活性、肝功能和肝组织学变化。结果 各组1w生存率均为100％;30％POLT组术后2d达到增殖高峰,峰值与PH组无差异（P〉0．05）;其肝功能酶学指标在术后1、3d明显升高,组织学见肝细胞核分裂极其活跃。结论 冷缺血1h的30％供肝和肝切除后肝脏具有同样的增殖活性,仅增殖高峰稍晚;较短的冷缺血时间以及熟练的手术技术可能对小体积供肝的再生起重要作用。     

Liver fatty acid-binding protein: an early and sensitive plasma marker of hepatocellular damage and a reliable predictor of graft viability after liver transplantation from non-heart-beating donors

OBJECTIVE: Liver fatty acid-binding protein (L-FABP) is a small protein (15 kD) involved in the intracellular transport of long-chain fatty acids in the liver. The L-FABP is regarded as a sensitive marker for liver cell damage. In a pig model for liver transplantation (LTx) from non-heart-beating donors (NHBD), we evaluated plasma changes of L-FABP early after reperfusion of grafts exposed to increasing periods of warm ischemia (WI). METHODS: Porcine livers were procured after 0, 15, 30, 45, and 60 minutes' WI. After 4 hours' cold ischemia (CI), LTx was performed. Primary graft nonfunction (PNF) and day 4 survival were recorded. Plasma samples were collected prior to and 15, 60, and 180 minutes after graft reperfusion for determination of L-FABP and aspartate transaminase (AST). RESULTS: Early after reperfusion, levels of L-FABP correlated well with the duration of WI. The PNF developed in 100% of animals after 60 minutes of WI, 50% after 30, and 45 minutes' WI, and was absent after no WI and 15 minutes of WI. Day 4 survival was 100% in 0 minutes' WI, 83% in 15 minutes' WI, 50% in 30 and 45 minutes' WI, and 0% in 60 minutes of WI. CONCLUSIONS: Plasma levels of L-FABP correlated well with WI and concomitant hepatocellular damage in LTx from NHBD. Monitoring of posttransplant L-FABP plasma levels is a valuable new tool to quantify early the extent of parenchymal cell damage of NHBD livers and to predict their viability and function.     

Effects of deletion variant of hepatocyte growth factor on reduced-size liver transplantation in rats.

BACKGROUND: The deletion variant of hepatocyte growth factor (dHGF) exerts mitogenic and antifibrotic effects. The purpose of this study was to evaluate the effect of dHGF on rats that had undergone syngeneic or allogeneic reduced-size (60%) orthotopic liver transplantation (ROLT). METHODS: Starting immediately after the syngeneic (Lewis to Lewis) and allogeneic (Lewis to Brown Norway) ROLT, 500 microg/kg dHGF was administered i.v. twice a day until the day the rats were killed. Its effect on hepatic graft weight, regeneration, and biochemical parameters was evaluated. RESULTS: dHGF promoted restoration of the liver volume and liver regeneration as well as protein synthesis in the rats that underwent syngeneic ROLT. In the rats that underwent allogeneic ROLT, dHGF reduced the level of serum cytosolic enzymes related to acute cellular rejection, but a significant improvement in liver regeneration and protein synthesis was not seen. When tacrolimus was administered to prevent rejection of the allogeneic grafts, the beneficial effect of dHGF was apparent, and was as beneficial as in syngeneic ROLT. CONCLUSIONS: Administering dHGF after liver transplantation augments the regeneration and functional recovery of partial liver grafts and reduces hepatocyte injury in acute cellular rejection.     

大鼠心脏停搏供体肝移植的实验研究

目的 探讨大鼠原位肝移植过程中,供肝可能耐受心脏停搏热缺血损伤的时间极限,方法 雄性SD大鼠,以供肝获取前供体大鼠经历心脏停搏时间0、15、30、45、60min分为5组（HB组、N-15组、N-30组、N-45组和N-60组）,而后行大鼠原位肝移植,比较各组的术后肝功能、肝脏病理和存活率。结果 HB、N-15、N-30、N-45和N-60的大鼠肝移植术后1周存活率分别为：100%（8/8）、75%（6/8）、62.5%(5/8)、25%（2/8）和0%（0/8）。其中HB、N-15、N-30组和N-45组的大鼠肝移植最长存活时间超过30d。结论 大鼠供肝对于热缺血损伤的时间应越短越好,但耐受30min心脏停搏热缺血损伤时,仍可使1周存活率达62.5%,极限时间为45min。     

无细胞的异体神经修复鼠坐骨神经缺损

目的 通过化学萃取同种异体神经,去除髓鞘和雪旺细胞,形成无细胞基膜管后桥接鼠坐骨神经缺损,研究神经再生效果。方法 正常鼠坐骨神经用非变性生物剂处理后得到无细胞的基膜管,桥接鼠坐骨神经20mm缺损。实验分3组:无细胞基膜管移植组(A组),自体神经移植组(B组)和异体神经移植组(C组)。术后进行肌电图、光镜、电镜及图象分析仪检查。结果 A组再生神经有大量轴突通过移植体,术后2个月电生理检测再生神经的潜伏期及波幅低于B组(P<0.05),术后3个月2组差异无显著意义。髓鞘厚度在术后3个月时亦低于B组,差异有显著意义(P<0.05)。轴突直径及数目两组无差异。C组因无神经再生,结果无法测量。结论 这种无细胞基膜管移植体能支持轴突的生长和雪旺细胞的迁移,是一种良好的神经移植替代材料。     

In situ lung perfusion is a valuable tool to assess lungs from donation after circulatory death donors category I–II

Donations after circulatory death (DCD) lung grafts are an alternative to extend the donor pool in lung transplantation. This study investigates the use of an in situ lung perfusion system (ISLP) in the donor to evaluate category I–II lungs. Pigs were sacrificed by ventricular fibrillation. All animals underwent 20 min of cardiopulmonary resuscitation and 5 min hands‐off period after which heparin was administered. In group [WI‐1], this was followed by 1 h of warm ischemia (WI) and 2 h of topical cooling (TC). In group [WI‐2], 2 h of WI was followed by 1 h of TC. In group [WI‐0], there was a minimal period of WI and no TC. In all three groups, the lungs were then evaluated during 60 min with ISLP. [WI‐0] lungs showed a significantly higher compliance and Δ PO₂/FiO₂ compared with [WI‐1] and [WI‐2]. PaCO₂ and lactate production were higher in [WI‐2] versus [WI‐0]. Wet/Dry weight ratio was significantly higher in [WI‐2] compared with [WI‐0] in two lung biopsy locations. A high W/D weight ratio was correlated with a lower compliance, higher lactate production, and a higher PaCO₂. ISLP is an effective way to assess the quality of lungs from category I–II DCD donors.     

The significance of preserving the energy status and microcirculation in liver grafts from non-heart-beating donor

To complete a successful liver transplantation (LTx) from non-heart-beating donors (NHBD), it is necessary to both improve the energy status in liver grafts and to reduce the exposure to free radicals. This study investigated the effects of short perfusion with oxygenated buffer on the grafts prior to cold preservation. In addition, the effects of the antioxidant, biliverdin, for reduction of free radicals was investigated. Male Wistar rats were used. Livers were retrieved, preserved in UW solution, and perfused for 60 min with oxygenated Krebs-Henseleit solution. Rats were allocated to six groups as follows (n=5): (i) control group-no warm ischemia (WI) and cold preservation, (ii) HBD group--no WI with cold preservation for 6 h; (iii) NHBD group--with 30 min of WI and cold preservation, (iv) NM group--with WI including nafamostat mesilate infusion before cardiac arrest and cold preservation; (v) PRE group--with WI, 30-min pre-cold preservation perfusion with oxygenated buffer after cardiac arrest, and cold preservation, (vi) BV group-with the same treatment as the PRE group plus the addition of biliverdin to the pre-cold preservation perfusion. The portal flow volume, bile production, AST, and TNF-alpha in perfusate, energy charge (EC), and ATP level in the tissue, and histological findings were investigated. The portal flow volume in the NM, PRE, and BV groups were higher than in the NHBD group. The bile production in the PRE and BV groups were also higher than in the NHBD group. The EC and ATP level of the BV group after reperfusion were higher than those of the NHBD group. Pre-cold preservation perfusion and addition of biliverdin to perfusate improved viability of grafts from NHBD. The results indicate that the preservation of the energy status and microcirculation of the graft is important for successful LTx from NHBD.     

大鼠肝移植急性排斥反应中肿瘤坏死因子受体配体的表达及其意义

目的 观察肝移植排斥反应中移植肝脏内糖皮质激素诱导的肿瘤坏死因子受体配体(GITRL)的表达.方法 采用Kamada's二袖套法建立从Lewis到Brown Norway(BN)大鼠的肝移植排斥模型为排斥组(n=5),从BN到BN的肝移植模型为耐受组(n=5).术后24h,抽取血液,取肝脏及分离库普弗(Kupffer)细胞,检测肝脏上GITRL及肿瘤坏死因子(TNF)-α的表达,Kupffer细胞上GITRL的表达,检测血清及细胞上清液中TNF-α的表达.免疫组织化学染色强度采用Image-Pro Plus 6.0图像分析软件分析.结果 免疫耐受组和排斥组肝脏内的CITRL的平均染色强度分别为0.113±0.007和0.270±0.018(P＜0.05),TNF-α平均染色强度分别为0.114±0.004和0.141±0.005(P＜0.05),耐受组和排斥组的Kupffer细胞GITRL平均染色强度分别为0.206±0.017和0.337±0.018(P＜0.05),Kupffer细胞的培养上清液中,耐受组和排斥组TNF-α的值分别为(68.66±21.12)、(178.33±29.39)ng/L(P＜0.05).结论 在排斥的早期阶段肝脏及Kupffer细胞的GITRL表达增高,监测和干扰GITRL可能有益于肝移植急性排斥反应的早期诊断和处理.

Abstract：

Objective To investigate te changes of glucocorticoid induced tumor necrosis factor receptor ligand (GITRL) in hepatic allograft rejection. Methods Liver transplantation from Lewis rats (n = 5 ) to Brown Norway (BN) rats was performed by Kamada' s two-cuff technique as acute rejection group. Liver transplantation from BN to BN rats ( n = 5 ) was performed as tolerance group. Recipients were sacrificed at 24th h postoperation. Blood samples were collected and grafts were harvested, then Kupffer cells were isolated. GITRL and tumor necrosis factor (TNF)-α protein expression in the hver was tested by immunohistochemistry, and the GITRL expression in Kupffer cells by immunocytochemistry. Enzyme linked immunosorbent assay (ELISA) was employed to detect the changes of TNF-α protein in the serum and supernatant. The staining intensity was analyzed by Image-Pro Plus 6. 0 image analysis software. Results At 24th h postoperation, the liver GITRL expression levels in tolerance and rejection groups were 0. 113 ± 0. 007 and 0. 270 ±0. 018, respectively (P ＜0. 05). The TNF-α expression levels in the liver in tolerance and rejection groups were 0. 114 ± 0. 004 and 0. 141 ± 0. 005 respectively ( P ＜ 0.05 ). The GITRL expression levels in Kupffer cells in tolerance and rejection groups were 0. 206 ±0. 017 and 0. 337 ±0. 018 respectively (P ＜0. 05 ). As compared with tolerance group (68. 66 ±21.12) ng/L, TNF-α protein expression levels were up-regulated in the supernatant of rejection group ( 178.33 ± 29. 39 ) ng/L ( P ＜ 0. 05 ).Conclusion The expression of GITRL in the liver and Kupffer cells was increased in the early stage of rejection, and monitoring and interfering GITRL may be useful for the early diagnosis and management of an acute rejection in liver transplantation.     

肝移植时供肝耐受无心跳热缺血的安全时限实验研究

目的 探讨肝移植时供肝耐受无心跳热缺血损伤的安全时限。方法 建立广西巴马小型猪原位肝移植动物模型并按移植前供肝经历心脏停搏时间0、30、45、60min分为4组，观测肝移植后各组一周存活率、肝功能、肝脏病理和肝脏能量代谢以及术中肝脏复流后微循环改变。结果上述各组术后一周存活率分别为：100％(5／5)、100％(5／5)、60％(3／5)、20％(1／5)。肝脏能量代谢等指标的改变在供肝无心跳热缺血时间30min前是可逆的，随心跳热缺血时间的延长远渐向不可逆演变。结论 在本实验条件下，巴马小型猪心脏停搏供体肝移植时供肝耐受无心跳热缺血损伤的安全时限约为30min。